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Introduction: The increasing prevalence of infertility in male dogs in clinical practice mirrors current trends seen in human medicine. Acquired infertility is notably more common in dogs compared to congenital causes, with conditions such as testicular degeneration leading to irreversible loss of fertility. Current diagnostic methods for testicular degeneration, such as histopathological and cytological examinations, rely on testicular biopsy or fine needle aspiration, making them less feasible for routine use. Anti-Müllerian hormone (AMH), produced by Sertoli cells, has emerged as a potential alternative biomarker for testicular health, which can be measured in serum. This study evaluates AMH as a potential marker for testicular degeneration, using cryptorchid dogs as models for impaired fertility and altered testicular histology. Methods: The relationship between serum AMH levels and AMH tissue expression with impaired spermatogenesis and altered histology was investigated. Serum AMH levels were determined in intact, cryptorchid, and castrated individuals using an immuno-enzymatic ELISA kit and compared between subgroups based on testicular location. Tissue AMH immuno-expression was differentially quantified in two regions of interest (ROIs), the interstitial space and the seminiferous tubule, in both descended and retained gonads. Furthermore, testicles were analyzed using histomorphometric analysis in seminiferous tubules, while spermatogenesis was evaluated using the Johnsen score. Results: Serum AMH levels were positively correlated with AMH expression assessed in both interstitial space (ρ = 0.494, p ≤ 0.01) and seminiferous tubules (ρ = 0.610, p ≤ 0.001). Conversely, serum AMH levels showed a negative correlation with the seminiferous tubule area (ρ = -0.435, p ≤ 0.05). Smaller seminiferous tubule areas were linked to increased AMH reactivity in both seminiferous tubules (ρ = -0.774, p ≤ 0.001) and interstitial space (ρ = -0.725, p ≤ 0.001). Additionally, lower Johnsen scores were associated with higher serum AMH levels (ρ = -0.537, p ≤ 0.01) and elevated AMH expression in both seminiferous tubules (ρ = -0.756, p ≤ 0.001) and interstitial space (ρ = -0.679, p ≤ 0.001). Discussion: Our results suggest that higher serum levels and tissue expression of AMH are linked to smaller seminiferous tubules and poorer Johnsen scores, reflecting degenerative changes and Sertoli cell dysfunction in retained testicles. Given the similarities in the mechanisms that increase AMH levels in both cryptorchid and non-cryptorchid testicles affected by testicular degeneration, this study recommends using AMH as a marker for diagnosing testicular degeneration in dogs. [ABSTRACT FROM AUTHOR]

Background Objectives Materials and methods Results Conclusions Imatinib and dasatinib are tyrosine kinase inhibitors (TKIs) increasingly used to treat several diseases in both children and adults at fertile age. We have previously shown that imatinib has adverse effects on developing testis, and imatinib‐treated male patients have been reported to have reduced sperm counts. However, the cellular level effects of imatinib and dasatinib on adult male germ cells and germline stem cells (mGSCs) have not been thoroughly investigated.To analyze whether imatinib or dasatinib exposure ex vivo and in vitro is harmful to adult male rodent germ cells and mGSCs.Seminiferous tubule segments of adult male mouse or rat were cultured in the presence or the absence of imatinib or dasatinib. Stage‐specific effects were monitored by 3H‐thymidine incorporation assay (DNA synthesis), immunohistochemistry (cleaved Caspase‐3; apoptosis), immunofluorescence (KI67, GFRα1, STRA8, c‐KIT, LIN28A; proliferation and spermatogonial differentiation) and flow cytometry (Hoechst). Mouse mGSCs were exposed to imatinib and dasatinib to study proliferation, apoptosis, and differentiation.Imatinib decreased stage‐specific DNA synthesis, and induced apoptosis in cultured rat seminiferous tubule segments. Imatinib also had an adverse effect on mGSC proliferation both in vitro and ex vivo, but did not induce cell death in cultured mGSCs. Imatinib did not impinge on induction of spermatogonial differentiation but suppressed c‐KIT expression in nascent differentiating spermatogonia, providing a plausible mechanism for its pro‐apoptotic function in spermatogenic cells. Clinically relevant doses of dasatinib did not induce apoptosis in seminiferous tubules but decreased mGSC colony growth in vitro.Imatinib exposure ex vivo and in vitro impinges on male rodent germ cell proliferation and survival. The plausible mechanism in spermatogenic cells is the inhibition of SCF/c‐KIT signaling, and reduced expression of c‐KIT. Dasatinib did not show significant adverse effects with clinical doses ex vivo but inhibited mGSC colony growth in vitro. [ABSTRACT FROM AUTHOR]

Background Objectives Materials and Methods Results Discussion Conclusion Histological analysis of the testicular sections is paramount in infertility research but tedious and often requires months of training and practice.Establish an expeditious histopathological analysis of mutant mice testicular sections stained with commonly available hematoxylin and eosin (H&E) via enhanced deep learning modelAutomated segmentation and cellular composition analysis on the testes of six mouse reproductive mutants of key reproductive gene family, DAZ and PUMILIO gene family via H&E‐stained mouse testicular sections.We improved the deep learning model with human interaction to achieve better pixel accuracy and reduced annotation time for histologists; revealed distinctive cell composition features consistent with previously published phenotypes for four mutants and novel spermatogenic defects in two newly generated mutants; established a fast spermatogenic defect detection protocol for quantitative and qualitative assessment of testicular defects within 2.5–3 h, requiring as few as 8 H&E‐stained testis sections; uncovered novel defects in AcDKO and a meiotic arrest defect in HDBKO, supporting the synergistic interaction of Sertoli Pum1 and Pum2 as well as redundant meiotic function of Dazl and Boule.Our testicular compositional analysis not only could reveal spermatogenic defects from staged seminiferous tubules but also from unstaged seminiferous tubule sections.Our SCSD‐Net model offers a rapid protocol for detecting reproductive defects from H&E‐stained testicular sections in as few as 3 h, providing both quantitative and qualitative assessments of spermatogenic defects. Our analysis uncovered evidence supporting the synergistic interaction of Sertoli PUM1 and PUM2 in maintaining average testis size, and redundant roles of DAZ family proteins DAZL and BOULE in meiosis. [ABSTRACT FROM AUTHOR]

Organoids play pivotal roles in uncovering the molecular mechanisms underlying organogenesis, intercellular communication, and high-throughput drug screening. Testicular organoids are essential for exploring the genetic and epigenetic regulation of spermatogenesis in vivo and the treatment of male infertility. However, the formation of testicular organoids with full spermatogenesis has not yet been achieved. In this study, neonatal mouse testicular cells were isolated by two-step enzymatic digestion, and they were combined with Matrigel and transplanted subcutaneously into nude mice. Histological examination (H&E) staining and immunohistochemistry revealed that cell grafts assembled to form seminiferous tubules that contained spermatogonial stem cells (SSCs) and Sertoli cells, as illustrated by the co-expression of PLZF (a hallmark for SSCs) and SOX9 (a marker for Sertoli cells) as well as the co-expression of UCHL1 (a hallmark for SSCs) and SOX9, after 8 weeks of transplantation. At 10 weeks of transplantation, SSCs could proliferate and differentiate into spermatocytes as evidenced by the expression of PCNA, Ki67, c-Kit, SYCP3, γ-HA2X, and MLH1. Notably, testicular organoids were seen, and spermatids were observed within the lumen of testicular organoids after 16 weeks of transplantation, as shown by the presence of TNP1 and ACROSIN (hallmarks for spermatids). Collectively, these results implicate that we successfully established testicular organoids with spermatogenesis in vivo. This study thus provides an excellent platform for unveiling the mechanisms underlying mammalian spermatogenesis, and it might offer valuable male gametes for treating male infertility. [ABSTRACT FROM AUTHOR]

Background Objective Materials and methods Results Conclusions The PWWP domain‐containing proteins are involved in chromatin‐associated biological processes, including transcriptional regulation and DNA repair, and most of them are significant for gametogenesis and early embryonic development in mammals. PWWP3A, one of the PWWP domain proteins, is a reader of H3K36me2/H3K36me3 and a response factor to DNA damage. However, the physiological role of PWWP3A in spermatogenesis and fertility remains unclear.The goal of this study was to explore the function and mechanism of PWWP3A in the process of spermatogenesis.We generated V5‐Pwwp3a KI mice and PWWP3A polyclonal antibody to observe the localization of PWWP3A in vivo. Meanwhile, Pwwp3a KO mice was used to explore the function in spermatogenesis.We reported that PWWP3A is a predominant expression in the testis of mice. During spermatogenesis, PWWP3A exhibits the temporal expression from early‐pachytene to the round spermatids. The results of spermatocyte spreading and immunostaining showed that PWWP3A aggregated on the XY body, which then diffused as the XY chromosome separated at late‐diplotene. Although the depletion of PWWP3A had no obvious reproductive defects in young male mice, there were observed morphological abnormalities in sperm heads. Immunoprecipitation demonstrated the interaction of PWWP3A with DNA repair proteins SMC5/6; however, PWWP3A deficiency did not result in any meiotic defects. Notably, the testes of aged male Pwwp3a KO mice displayed pronounced degeneration, and were characterized by the presence of vacuolated seminiferous tubules. Furthermore, RNA‐seq analysis revealed an upregulation in the expression of genes which may be involving in immunoregulatory and inflammatory response pathways in aged Pwwp3a KO mice with testicular degeneration.Our study showed that PWWP3A was highly enriched in the mouse testis, and the Pwwp3a KO mice were fertile. However, the aged Pwwp3a KO male mice displayed testicular atrophy that may be due to changes in the immune micro‐environment or abnormal repair of DNA damage. [ABSTRACT FROM AUTHOR]

Simple Summary: The aim of this experiment was to investigate the distinct effects of microplastic-polyvinyl chloride (PVC) on the reproductive system, intestinal tissue structure, and microbial flora in male and female mice. The impact of PVC microplastics on intestinal microflora differed between males and females, showing contrasting trends. Both male and female mice exhibited impaired structural integrity of the intestine and reproductive systems due to polyvinyl chloride microplastics. Additionally, there was a significant correlation between disturbances in intestinal microflora caused by PVC microplastics and reproductive system function. The pervasive use of plastics in numerous industrial sectors has resulted in the circulation of microplastics across diverse ecosystems and food chains, giving rise to mounting concerns regarding their potential adverse impacts on biological systems and the environment. The objective of this experiment was to investigate the distinct effects of microplastic-polyvinyl chloride (PVC) exposure on the reproductive system, intestinal tissue structure, and intestinal microbial flora of both male and female mice. A total of 24 4-week-old Kunming mice were randomly assigned to one of four groups: male control group (CM), female control group (CF), male PVC test group (PVCM), and female PVC test group (PVCF) (n = 6). The findings revealed that in terms of the reproductive system, the PVCM group exhibited an impaired testicular structure with an irregular arrangement and a significant reduction in spermatogonia, spermatocytes, and spermatozoa within the seminiferous tubules (p < 0.01). The PVCF group exhibited a notable decrease in ovarian follicles (p < 0.01), accompanied by a reduction in uterus volume, fallopian tube volume, and muscle layer thickness, all of which also decreased significantly (p < 0.01). In comparison to the control groups, exposure to PVC resulted in a reduction in the width and height of the intestinal villi, accompanied by an increase in crypt depth. This led to a significant alteration in the ratio of villus height to crypt depth (V/C) (p < 0.01). Moreover, a reduction in microbial species diversity was observed within both the PVCM and PVCF groups; additionally, it was accompanied by contrasting changes in relative abundance and functional gene profiles among the major intestinal flora constituents. In summary, the findings indicate that PVC induces damage to both male and female mice reproductive and digestive systems, further exhibiting notable sex-dependent effects on mouse intestinal microflora composition, which correlates significantly with its impact on reproductive organs. [ABSTRACT FROM AUTHOR]

Amiodarone hydrochloride is an antiarrhythmic agent that is widely prescribed. However, it has serious side effects that approximately affect the whole body organs. In our study, we aimed to assess the possible effects of chronic administration of two different doses of amiodarone hydrochloride on the oxidative and inflammatory parameters as well as the histological morphology and ultrastructure of the seminiferous tubules of adult male albino rats. Forty rats were divided into four groups; Control group 1: each rat did not receive any drugs at all. Control group 2: each rat received 3 ml of 0.16% methylcellulose, orally and daily for 4 weeks. Low dose amiodarone group: each rat received 3 ml of 0.16% methylcellulose contained 3.6 mg amiodarone, orally and daily for 4 weeks. High dose amiodarone group: each rat received 3 ml of 0.16% methylcellulose contained 7.2 mg amiodarone, orally and daily for 4 weeks. Blood samples were collected for measuring serum levels of malondialdehyde, superoxide dismutase, interleukin-6 and tumor necrosis factor-alpha. Testes specimens were examined to assess the morphological changes and the level of expression of caspase-3 apoptotic marker. The results indicated that; amiodarone hydrochloride could induce a dose-dependent toxicity, causing oxidative stress, inflammation, cellular degeneration, deposition of collagen and enhanced apoptosis in the seminiferous tubules. [ABSTRACT FROM AUTHOR]

Background Objectives Materials and Methods Results Discussion and Conclusion Testicular torsion/detorsion can cause testis loss and infertility. Aloperine is a major active alkaloid extracted from Sophora alopecuroides Linn. It has been shown to have organ‐protective effects. However, the effects of aloperine on the testis and its underlying mechanisms remain unclear.This study investigated the effect of aloperine on testicular torsion/detorsion injury in rats.Male Sprague‐Dawley rats were randomized to the sham‐operated (sham), testicular I/R (TI/R), or aloperine preconditioning (ALOPre) or postconditioning (ALOPost) groups. All rats except for the sham‐operated rats were subjected to 3 h of right spermatic cord torsion (720°, clockwise), followed by 3 h of detorsion. Aloperine (10 mg/kg) was intravenously administered before testicular torsion (ALOPre) or at the onset of testicular detorsion (ALOPost). The therapeutic efficacy of aloperine was evaluated by histological analysis, oxidative stress evaluation, inflammatory response examination, apoptosis analysis, protein analysis, and immunohistological assessment.Compared with TI/R, aloperine protected both the ipsilateral and contralateral testes against unilateral testicular I/R, as evidenced by a reduced testicular weight to body weight (TW/BW) ratio (ALOPre: p = 0.0037; ALOPost: p = 0.0021) and volume (ALOPre: p = 0.0020; ALOPost: p = 0.0009), less structural damage with better Johnsen (ALOPre: p = 0.0013; ALOPost: p = 0.0021), and Cosentino scores (ALOPre: p < 0.0001; ALOPost: p < 0.0001), increased mean seminiferous tubule diameter and mean seminiferous tubule epithelial height, decreased testicular apoptosis, and less oxidative stress and inflammatory response. In addition, aloperine significantly stimulated the phosphorylation of signal transducer and activator of transcription (STAT)‐3 in the ipsilateral testes following detorsion. Administration of Ag490 suppressed STAT‐3 phosphorylation, thereby abrogating the protective effects exerted by aloperine on the ipsilateral testis.Aloperine has a strong testicular protective effect on the ipsilateral and contralateral testes after testicular torsion/detorsion. This aloperine‐induced ipsilateral testicular protection is mediated via the STAT‐3 signaling pathway. [ABSTRACT FROM AUTHOR]

Background: Differences of sex development (DSD) are congenital conditions in which chromosomal, gonadal, or phenotypic sex is atypical. In more than 50% of human DSD cases, a molecular diagnosis is not available. In intensively farmed pig populations, the incidence of XX DSD pigs is relatively high, leading to economic losses for pig breeders. Interestingly, in the majority of 38, XX DSD pigs, gonads still develop into testis-like structures or ovotestes despite the absence of the testis-determining gene (SRY). However, the current understanding of the molecular background of XX DSD pigs remains limited. Methods: Anatomical and histological characteristics of XX DSD pigs were analysed using necropsy and HE staining. We employed whole-genome sequencing (WGS) with 10× Genomics technology and used de novo assembly methodology to study normal female and XX DSD pigs. Finally, the identified variants were validated in 32 XX DSD pigs, and the expression levels of the candidate variants in the gonads of XX DSD pigs were further examined. Results: XX DSD pigs are characterised by the intersex reproductive organs and the absence of germ cells in the seminiferous tubules of the gonads. We identified 4,950 single-nucleotide polymorphisms (SNPs) from non-synonymous mutations in XX DSD pigs. Cohort validation results highlighted two specific SNPs, "c.218T > C" in the "Interferon-induced transmembrane protein 1 gene (IFITM1)" and "c.1043C > G" in the "Newborn ovary homeobox gene (NOBOX)", which were found exclusively in XX DSD pigs. Moreover, we verified 14 candidate structural variants (SVs) from 1,474 SVs, identifying a 70 bp deletion fragment in intron 5 of the WW domain-containing oxidoreductase gene (WWOX) in 62.5% of XX DSD pigs. The expression levels of these three candidate genes in the gonads of XX DSD pigs were significantly different from those of normal female pigs. Conclusion: The nucleotide changes of IFITM1 (c.218T > C), NOBOX (c.1043 C > G), and a 70 bp deletion fragment of the WWOX were the most dominant variants among XX DSD pigs. This study provides a theoretical basis for better understanding the molecular background of XX DSD pigs. Plain language summary: DSD are conditions affecting development of the gonads or genitalia. These disorders can happen in many different types of animals, including pigs, goats, dogs, and people. In people, DSD happens in about 0.02–0.13% of births, and in pigs, the rate is between 0.08% and 0.75%. Pigs have a common type of DSD where the animal has female chromosomes (38, XX) but no SRY gene, which is usually found on the Y chromosome in males. XX DSD pigs may look like both males and females on the outside and have testis-like or ovotestis (a mix of ovary and testis) gonads inside. XX DSD pigs often lead to not being able to have piglets, slower growth, lower chance of survival, and poorer meat quality. Here, we used a method called whole-genome de novo sequencing to look for variants in the DNA of XX DSD pigs. We then checked these differences in a larger group of pigs. Our results reveal the nucleotide changes in IFITM1 (c.218T > C), NOBOX (c.1043 C > G), and a 70 bp deletion fragment in intron 5 of the WWOX, all linked to XX DSD pigs. The expression levels of these three genes were also different in the gonads of XX DSD pigs compared to normal female pigs. These variants are expected to serve as valuable molecular markers for XX DSD pigs. Because pigs are a lot like humans in their genes, physiology, and body structure, this research could help us learn more about what causes DSD in people. Highlights: XX DSD pigs have abnormal development of reproductive organs showing both male and female characteristics and lack of germ cells in the seminiferous tubules. Whole-genome de novo sequencing was used to reveal the characteristics of genomic variants in XX DSD pigs. The nucleotide changes of IFITM1 (c.218T > C), NOBOX (c.1043 C > G), and a 70 bp deletion fragment of WWOX were the dominant variants in XX DSD pigs, and these three genes were significantly changed in the gonadal expression levels of the XX DSD pigs compared to those of normal females. [ABSTRACT FROM AUTHOR]

Overveiw: Cisplatin (CIS) is a perfect antineoplastic medicine resulting in severe testicular toxicity. Melatonin is a potent antioxidant that can prevent gonadal toxicity. exosomes have been demonstrated to have a vital effect in tissue regeneration. Aim to the Work: To evaluate the potential protective effects of melatonin and exosomes developed from bone marrow mesenchymal stem cells (BMSC-EX) in testicular toxicity caused by cisplatin. Materials and Methods: Fifty rats were divided in a random way into five equal groups. Group I (control group). Group II were given a single dose (8 mg/kg) of cisplatin intraperitoneally I.P. Group III (melatonin group): intra peritoneal administration of melatonin (4 mg/kg) started 5 days before cisplatin injection for 15 days. Group IV (exosomes group): a single intra venous dose of 100 μg BMSC-EX one day before cisplatin dose. Group V (melatonin and exosomes). Specimens of the testicles were collected and processed for immunohistochemical and histological analysis. Results: Group II declared distortion, irregular outline of the seminiferous tubule, with critical accumulation (P <0.01) of collagen fibers. A substantial elevation was present (P <0.01) in TNF-α immunostaining and substantial reduction (P <0.01) was seen in AR and PCNA immunostaining in comparison to control group. Groups III and IV exhibited some microscopic histological variations development, a critical reduction (P <0.01) was also observed in collagen fibers deposition and TNF-α immunostaining and critical elevation (P <0.01) was present in AR and PCNA immunostaining in comparison to group II. In comparison to control group, group V histological structure was similar. Conclusion: Each of melatonin and exosomes can protect against testicular toxicity caused by cisplatin when administered before and during cisplatin therapy, but their combined administration has better results. [ABSTRACT FROM AUTHOR]

Objective: Cyclophosphamide (CP) is an alkylating agent commonly used in cancer treatment. It is known to have detrimental effects on the reproductive system, including the potential to cause infertility. Recently, herbal remedies have gained traction as a complementary approach to addressing these side effects. In this study, our goal was to investigate whether the aqueous-alcoholic extract of Withania somnifera (WS) could mitigate the adverse impacts of CP on testicular tissue. Methods: Animals were randomly assigned to one of the following groups: control, WS (500 mg/kg), CP (100 mg/kg), CP+WS pre-treatment, and CP+WS post-treatment. WS was administered orally through gavage for 1 month. We assessed sperm parameters, testicular histopathology, and the expression of the Bax and Bcl2 genes in the experimental groups. Results: Sperm parameters (including count, viability, and motility), the number of spermatogonia, the seminiferous tubule diameter, and Bcl2 gene expression, significantly decreased after CP injection (p<0.05). Conversely, the number of immotile sperm and Bax gene expression significantly increased (p<0.05). Treatment with WS, especially when administered as a pre-treatment, ameliorated the sperm parameters, histological alterations, and the expression of apoptosis-related genes (p<0.05). Conclusion: The data suggest that WS may mitigate the detrimental effects of CP on testicular tissue by reducing apoptosis. Consequently, WS has the potential to be used as an adjunctive therapy to reduce the complications associated with CP treatment. [ABSTRACT FROM AUTHOR]

Studies have shown an association between homocysteine (Hcy) and several pathological conditions, including cardiovascular disease. However, the relationship between Hcy and testicular disorders is still unclear. The present study investigated the effects of Hcy on testicular structure. Thirteen adult Wistar albino rats were divided into two groups. The control group and the methionine group received an oral solution of L-methionine 200 mg /kg per day for six months. Blood samples and testes were collected for subsequent experiments. Serum levels of Hcy and testosterone were analyzed. We examined the histopathological and immunohistochemical localization of GATA-4, vimentin (VIM), connexin-43 (Cx-43), occludin (OCLN), androgen receptor (AR), estrogen receptor 1 (ESR1), estrogen receptor 2 (ESR2) in the rat testis. Results showed that rats receiving methionine showed a reduction in body and testicular weights. Histological and immunohistochemical examination of the testicular tissues of methionine group showed altered structure of seminiferous tubules with decreased immunoexpression of Cx-43, GATA-4, OCLN,VIM, AR, ESR1 and ESR2 compared to control. The results suggest that Hcy has a negative effect on the testes with a probable disruption of spermatogenesis. [ABSTRACT FROM AUTHOR]

N6-methyladenosine (m6A), an RNA post-transcriptional modification, plays a crucial role in spermatogenesis. Cattle-yaks are interspecific hybrid offsprings of yak and cattle, and male cattle-yaks are sterile. This study aims to investigate the role of m6A modification in male cattle-yak infertility. Herein, testicular tissues were analyzed via histological observations, immunohistochemical assays, reverse-transcription quantitative polymerase chain reaction, Western blotting, and immunofluorescence assays. The results revealed that male cattle-yaks presented smaller testes (5.933 ± 0.4885 cm vs. 7.150 ± 0.3937 cm), with only single cell layers in seminiferous tubules, and weakened signals of m6A regulators such as METTL14 (methyltransferase-like 14), ALKBH5 (alpha-ketoglutarate-dependent hydroxylase homolog 5), FTO (fat mass and obesity-associated protein), and YTHDF2 (YTH N6-methyladenosine RNA binding protein F2), both at the RNA and protein levels, compared with those of yaks. Altogether, these findings suggest that m6A modification may play a crucial role in male cattle-yak sterility, providing a basis for future studies. [ABSTRACT FROM AUTHOR]

Cypermethrin (CYP) is a synthetic pyrethroid. It acts as an endocrine disruptor which negatively affects animal's reproductive and developmental processes of progeny. So, The present study aimed to examine the toxicity of oral administration of CYP at three different doses (25mg, 50mg, and 75mg/kg) on the reproductive organs of albino rats and also on the growth and development of pups. For this study, animals were divided into four groups: Group I was the control group, and the remaining three were experimental groups that received CYP at a dose of 25mg (group II), 50mg (group III), and 75mg (group IV) per kg body weight (BW) of rat per day for 30 days. There are sixteen animals in each group-eight males and eight females. Histological studies of reproductive organs suggested that CYP exposure caused alteration in the structure of the ovary and testis tissue. In females, significant changes in the ovary, such as distorted follicles, loss of follicular stages, presence of antral follicles, damaged granulosa and theca cells, and blood vessel congestion, were observed, while in males, there were alterations in testes tissue damage in seminiferous tubules, reduced number of germinal cells, and reduction in the size of seminiferous tubules. A significant reduction was observed in the number, length, eyes opening, fur arrival time, and size of pups. Fertility was also recorded up to 80% in the 50 and 75mg CYP-treated female rats. The study suggested that CYP exposure caused gonadal toxicity in albino rats and affected the development of neonates. [ABSTRACT FROM AUTHOR]

This study investigated the effects of transplanted testicular stromal stem cells (tSSCs) on surgically damaged testis tissue. Ten‐week‐old male Wistar albino rats were divided into three groups: control (n = 6), damage (DG) (n = 6) and testicular stromal stem cell (TSSC) (n = 6) groups. Surgically induced damage was inflicted on the left testes of both the DG and TSSC groups, with no intervention on the right testes. In the TSSC group, damaged testes were treated with transplanted tSSCs, followed by orchiectomy after 15 days. Testes tissues were stained with haematoxylin–eosin (H&E), and recovery rates of functional structures were assessed by modified Johnsen scoring. The effects of tSSCs on testicular tissue were demonstrated by immunohistochemistry using BAX, BCL‐2 and caspase 3. Serum testosterone levels were analysed using the enzyme‐linked immunosorbent assay (ELISA) method. Surgical damage caused germ cell degeneration in some seminiferous tubules and a decrease in interstitial areas. With tSSC treatment, improvements in testicular architecture were identified through spermatogenesis in the seminiferous tubules and normal histological structures in the interstitial areas. Correspondingly, in the modified Johnsen score, the DG group showed a significant difference compared to the other groups (p = 0.001). High expressions of BAX, BCL‐2 and caspase‐3 in the DG group revealed prominent features of apoptosis. With the injection of tSSCs, these expressions significantly normalized according to H score analysis (all p = 0.004). Although serum testosterone levels in the tSSC group were higher compared to the control and DG groups, this difference was not statistically significant (p = 0.119). This study suggests transplanting tSSCs could accelerate tissue healing after testicular sperm extraction (TESE) surgery for azoospermia patients, potentially paving the way for a new and important clinical treatment. [ABSTRACT FROM AUTHOR]

The purpose of this study is to examine the effect of baicalin on cyclophosphamide (CP)-induced testicular damage in rats. Twenty-eight Wistar albino rats were assigned to the control, baicalin, CP, and CP+Baicalin groups. A single dose of CP (200 mg/kg) was induced intraperitoneally (i.p.) in the CP and CP+Baicalin groups. Baicalin (100 mg/kg/day, i.p.) was administered in the baicalin and CP+ Baicalin groups for 6 days. After sacrification, the testes, epididymis and blood samples were taken. Testis tissues were investigated for general morphology, proliferating, and apoptotic cells. Oxidative stress parameters in testis tissue and hormone levels in serum were examined by biochemical analysis. Sperm analysis was performed on smear samples obtained from the epididymis. In CP + Baicalin group, decreases in both the number of abnormal spermatozoa and degeneration of seminiferous tubules were observed compared to the CP group. Also, decreased apoptotic cells and increased proliferative cells were detected in these seminiferous tubules. Furthermore, the treatment with Baicalin reversed the changes observed in the markers of oxidative damage including malondialdehyde, total oxidant status, and oxidative stress index, and markers of the antioxidant defense system including glutathione, total antioxidant capacity, superoxide dismutase, and catalase except for glutathione s-transferase (GST) in testicular tissue. Decreased serum levels of follicle-stimulating hormone, luteinizing hormone, and testosterone related to CP significantly improved in CP + Baicalin group. Conclusion: Baicalin attenuates CP-induced testicular damage by inhibiting oxidative stress possibly due to its antioxidant properties. Considering on the proposed beneficial effects of Baicalin against testicular damage induced by CP, it might have the potential to be a promising agent for male infertility related to chemotherapy. [ABSTRACT FROM AUTHOR]

Clinical and epidemiological studies indicated that there is an increased incidence of male infertility, with many cases that do not respond well to treatments. This study is aimed at evaluating the therapeutic effects of verapamil and tamoxifen against infertility, which may provide an essential strategy for the treatment of this disorder. A casecontrol study was conducted on 64 male albino rats, divided into four groups (16 for each); group A (control group); group B (infertile group that is induced by the ornidazole); and groups C and D, which are pretreated with verapamil and tamoxifen, respectively, in combination with ornidazole every day for four weeks. Serum levels of testosterone, follicle-stimulating hormone (FSH), luteinizing hormone (LH), glutathione (GSH), nitric oxide (NO), malondialdehyde (MDA), and superoxide dismutase (SOD) were assessed, and sperm parameters, including sperm count, sperm motility percentage, sperm morphology percentage, and sperm viability, were investigated. Furthermore, Motic Image Plus 2.0 ML was used to measure the mean diameter of the seminiferous tubules (SNT), and a morphometric analysis of 20 tubules per rat was also performed. The results revealed that both verapamil and tamoxifen provide a significant improvement in sperm count, motility, morphology, and dead sperm, with a significant improvement in testosterone, FSH, LH, GSH, NO, MDA, and SOD levels compared with the infertile group. Histophotometric results showed that verapamil and tamoxifen improved the number and size of SNT, Leydig cells, and Sertoli cell counts, which may indicate the promise of using these agents in the treatment of male infertility. [ABSTRACT FROM AUTHOR]

Objective: The objective of this study was to assess the protective effects of Heracleum persicum L. leaves extract (HPE) against oxidative damage induced by gentamicin (GM) in the testes of rats through biochemical, histopathological, and immunohistochemical approaches. Materials and Methods: Thirty-six male Wistar rats were divided into six groups (n=6/group) for 50 days. On day 51, the study assessed serum levels of luteinizing hormone (LH), folliclestimulating hormone (FSH), and testosterone (TT), as well as antioxidant enzyme activity, nitric oxide levels, and various parameters related to testicular tissue (including the ferric reducing ability of plasma (FRAP), thiol, and thiobarbituric acid reactive substances (TBARS) levels). The stereological indices of seminiferous tubules were measured using serial sections of testicular tissue stained with hematoxylin and eosin, while the apoptosis rate of testicular parenchymal cells (p53, Caspase-3, and Bcl-2 positive cells) was also determined. Results: In the groups treated with HPE, particularly at 750 mg/kg, there was a significant increase (p<0.05) in LH, FSH, and TT hormone levels, an enhanced serum antioxidant enzyme activity and significantly reduced (p<0.05) nitric oxide levels. HPE inhibited the apoptotic pathway involving Bax/p53/Caspase-3 (significantly decreased (p<0.05) all three genes), thereby preserving the structure and function of the testicular tissue. Consequently, the number of p53 and Caspase-3 positive testicular cells decreased significantly (p<0.05), while the number of Bcl-2 positive cells increased. Conclusion: HPE demonstrated potential in protecting the function and structure of testis against toxic and oxidative damages. [ABSTRACT FROM AUTHOR]

Objective: The aim of the current study was to assess the effects of Nigella sativa essential oil on testicular toxicity in mice induced by dexamethasone. Materials and Methods: Forty NMRI mice were randomly divided into four groups. The first group (Sham) received 1 ml per day of normal saline by intraperitoneal (i.p.) injection for 7 days. The second group (Control) received (i.p) injection of 5 mg/kg dexamethasone for 7 days. The third group (Dexa+ N.S 5 mg/kg) received dexamethasone (5 mg/kg) and gavaged 5 mg/kg N. sativa essential oil for 7 days. The fourth group (N.S 5 mg/kg) for 7 days was gavaged 5 mg/kg N. sativa. Histopathology of testis, spermatogenesis, and sperm fertility rate were assessed. Results: The results of histopathology assessments showed that in the third group, all histopathology criteria were decreased compared to the second group. The number of seminiferous tubules that had abnormal spermatogenesis in Johnsen’s score was slightly decreased in the third group compared to the second group. Furthermore, in the third group, embryo formation criteria were increased. Conclusion: The data of this research demonstrate that N. sativa improves spermatogenesis defects and sperm fertility in mice treated with dexamethasone. [ABSTRACT FROM AUTHOR]

Levamisole is an anti-helminthic drug developed and introduced in veterinary medicine, and it has been used more frequently after the inclusion of its usage in human medicine regarding disorders with immunomodulatory properties. In recent years, it has started to attract attention since it has beneficial effects on the treatment of COVID-19 due to its immunomodulatory properties. To investigate the effects of levamisole on sexual behavior and the reproductive system in male rats, two groups were formed the vehicle (n = 10) and levamisole (n = 10) groups. The vehicle group was given purified water whereas the levamisole group was administered with levamisole (2 mg/kg) by oral gavage daily for 4 weeks. Levamisole treatment significantly increased the mount latency (ML, P < 0.001) as well as the intromission latency (IL, P < 0.01). It also significantly prolonged postejaculatory interval (PEI, P < 0.01), decreased copulatory rate (CR, P < 0.05), and sexual activity index (SAI, P < 0.05). It significantly decreased serum monoamine oxidase A (MAO-A) levels (P < 0.05). Additionally, levamisole induced disorganizations of germinal epithelial cells of seminiferous tubules, congestion, edema in the interstitial area, and metaphase arrest in some spermatocytes (P < 0.001), and it significantly increased the immunohistochemical expressions of apoptotic Bax and cytochrome c, which is crucial proapoptotic protein, in the testis (P < 0.001). Also, levamisole significantly upregulated the mRNA levels of the apoptosis-related key regulatory genes, including Bax (Bcl-2-associated X protein, P = 0.05) and Bax/Bcl-2 ratio (P < 0.01) in testis. The current research is the first to show that levamisole may decrease sexual performance, potency, sexual motivation, and libido and induce apoptosis in the testis. [ABSTRACT FROM AUTHOR]

Gender‐affirming hormone therapy (GAHT) can help transgender and/or gender diverse (TGD) individuals achieve emobidment goals that align with their transition needs. Clinical evidence from estradiol (E)‐GAHT patients indicate widespread changes in tissues sensitive to E and testosterone (T), particularly in the reproductive system. Notably, E‐GAHTs effects on hormones and reproduction vary greatly between patients. With the goal of informing clinical research and practice for TGD individuals taking E, this study examines intact male mice implanted with capsules containing one of three different E doses (low 1.25 mg; mid 2.5 mg; high 5 mg), or a blank control capsule. All E‐GAHT doses suppress T and follicle stimulating hormone levels while elevating E levels. Only the high E‐GAHT dose significantly supresses luteinizing hormone levels. All E‐GAHT doses affect epididymis tubule size similarly while seminiferous tubule morphology and bladder weight changes are dose‐dependent. E‐GAHT does not alter the presence of mature sperm, though E‐exposed sperm have altered motility. These data represent the first evidence that mouse models offer an effective tool to understand E‐GAHTs impact on reproductive health and the dose‐dependent effects of this model permit examinations of diverse patient outcomes. [ABSTRACT FROM AUTHOR]

Benzo[a]pyrene (BaP) is a contaminant that is generated in the environment through processes such as smoke, incomplete combustion of fossil fuels, vehicle exhaust emissions, entry into the body is through inhalation, and consumption of contaminated food. It is an omnipresent environmental pollutant with unavoidable exposure. BaP metabolites are observed in the male reproductive system, especially in the testes and epididymis of animals, and are responsible for reduced testicular and epididymal function. The protective effect of atorvastatin (ATV) on testicular damage was investigated previously. The aim of the present study was to investigate the protective effect of ATV on testicular toxicity induced by benzo[a]pyrene (BaP) during pregnancy in Wistar rats. This experimental laboratory study involved 40 adult rats, divided into seven groups and maintained under standard environmental conditions. The groups received different diets [control, corn oil, ATV (10 mg/kg), BaP (10 and 20 mg/kg), and ATV + BaP (10 and 20 mg/kg)] at gestation Days 7–16, orally. Male offspring were examined 10 weeks after birth. Testis and serum samples were collected, and testosterone level, malondialdehyde (MDA), and glutathione (GSH) were measured. Histological and immunohistochemical assays were performed under a light microscope. Statistical analysis was conducted using SPSS, with analysis of variance and Tukey tests to assess significant differences between groups. ATV significantly reduced MDA, a marker of lipid peroxidation and oxidative stress in rat testes following BaP administration. Treatment with ATV at doses of 10 mg/kg increased GSH levels, correcting disruptions in the antioxidant system caused by BaP. Testosterone concentration in rats treated with ATV and BaP substantially prevented the decrease induced by BaP. Histomorphometry revealed that ATV significantly prevented the detrimental effects of BaP on the thickness of spermatogenic epithelium and the diameter of seminiferous tubules. Under ATV treatment, testicular tissue histopathology improved, and spermatogenesis returned to a almost back to normal state. Caspase‐3 expression decreased, and apoptosis activity in testicular tissue improved under ATV treatment, indicating a positive effect of ATV in reducing apoptotic damage caused by BaP. In conclusion, exposure to BaP can induce oxidative stress‐related damage to testicular tissue, as evidenced by an increase in MDA levels, which ATV treatment can mitigate. Additionally, ATV enhances intracellular antioxidant GSH and protects the testes against BaP‐induced damage while increasing testosterone levels, which are reduced due to exposure to BaP. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is an environmental pollutant known as endocrine disruptor. Cd has been reported to induce perturbations of the testicular functions and the subsequent decline of the male fertility of both animals and humans. Chlorella vulgaris (ChV) a species of green microalga has been reported to have multiple beneficial activities such as anti-inflammatory, antioxidant, and antiapoptotic effects. Thus, this work was conducted to declare the benefits of Chlorella vulgaris (ChV) (500 mg/kg doses) against cadmium chloride CdCl2 (2 mg/kg doses) toxicity on the main and accessory reproductive organs' weight, structure, and function of male rats. Briefly, 40 adult male rats in 4 groups (n = 10) were used as follows; control, ChV, CdCl2, and CdCl2+ChV. (i) The 1st group was kept as control fed on pellet chow and water ad libitum. (ii) The second group is Chlorella vulgaris (ChV) group fed with C. vulgaris alga for 10 days (500 mg/kg BW). (iii) The third group was administrated CdCl2 (2mg/kg BW) via subcutaneous injection (S/C) daily for 10 days. (iv) The fourth group administered both CdCl2 and ChV with the abovementioned doses daily for successive 10 days. Our observations declared that cadmium exhibited an adverse influence on the testes and prostate gland architecture indicated by seminiferous tubule destruction, testicular edema, degeneration of Leydig cells, and prostate acini damage. All together affect the epididymal semen quality and quantity including sperm viability, motility, and count. Interestingly, ChV could restore the testicular architecture and spermatozoa regeneration accompanied by semen quality improvement and increased reproductive hormones including testosterone. On the other side, ChV suppresses reactive oxygen species (ROS) formation via enhancement the antioxidant-related genes in the testicular tissue including SOD, CAT, GSH, and MDA and maintaining spermatocyte survival via suppression of apoptotic related genes including caspase3 and activating steroidogenic related genes including StAR and HSD17β3 in the cadmium-treated testes. In this study, ChV could enhance male fertility under normal or stressful conditions and ameliorate the adverse effects of hazardous heavy metals that are widely distributed in our environment. [ABSTRACT FROM AUTHOR]

Serine protease 50 (PRSS50/TSP50) is highly expressed in spermatocytes. Our study investigated its role in testicular development and spermatogenesis. Initially, PRSS50 knockdown was observed to impair DNA synthesis in spermatocytes. To further explore this, we generated PRSS50 knockout (Prss50−/−) mice (Mus musculus), which exhibited abnormal spermatid nuclear compression and reduced male fertility. Furthermore, dysplastic seminiferous tubules and decreased sex hormones were observed in 4-week-old Prss50−/− mice, accompanied by meiotic progression defects and increased apoptosis of spermatogenic cells. Mechanistic analysis indicated that PRSS50 deletion resulted in increased phosphorylation of extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) and elevated levels of MAP kinase phosphatase 3 (MKP3), a specific ERK antagonist, potentially accounting for testicular dysplasia in adolescent Prss50−/− mice. Taken together, these findings suggest that PRSS50 plays an important role in testicular development and spermatogenesis, with the MKP3/ERK signaling pathway playing a significant role in this process. [ABSTRACT FROM AUTHOR]

Stem cell transplantation into seminiferous tubules of recipient testis could become a tool for fertility restoration, genetic improvement, or conservation of endangered species. Spermatogonial stem cells (SSCs) are primary candidates for transplantation; however, limited abundance, complexity for isolation and culture, and lack of specific markers have limited their use. Mesenchymal stromal/stem cells (MSCs) are multipotent progenitors that are simple to isolate and culture and possess specific markers for identification, and immune evasive and migratory capacities. The objective of the present study was to evaluate the potential for survival and colonization in seminiferous tubules of two different concentrations of bovine fetal adipose tissue-derived MSCs (AT-MSCs), native of pre-induced, and to compare the fate of bovine adult peripheral blood-derived MSCs (PB-MSCs) and SSCs after allogenic transplantation in testis of recipient bulls. In experiment 1, AT-MSCs at two concentrations (1x107 and 2x107; n = 3) or pre-exposed to 2 μM testosterone and 1 μM retinoic acid (RA) for 14 days (n = 5) were evaluated. In experiment 2, adult PB-MSCs and SSCs (4x107 cells each) pre-exposed to Sertoli cell conditioned media (SCs/CM; n = 4) for 14 days were compared. Each cell type was separately labelled with PKH26 and then transplanted into testes of 8-month-old recipient bulls. Four weeks (Exp. 1) and two weeks (Exp. 2) after transplantation, testicular tissue was processed for confocal microscopy detection of PKH26-positive cells. Mean number of PKH26-positive cells were higher (P < 0.05) in testis transplanted with 2x107 AT-MSCs in the proximal (6.7 ± 3.7) and medial (6.6 ± 3.2) sections compared to testis transplanted with 1x107 AT-MSCs (proximal: 1.9 ± 1; medial: 1.9 ± 1) sections or pre-induced AT-MSCs (proximal: 4.7 ± 5.6; medial: 3.8 ± 4.1). In Exp. 2, mean number of PKH26-positive SSCs in medial testicular section (22.5 ± 1.3) were higher (P < 0.05) compared to respective section in PB-MSCs group (17 ± 4.2). Thus, in vivo data indicates that a higher number of transplanted AT-MSCs resulted in more cells surviving and colonizing seminiferous tubules; however, pre-induction with testosterone and RA did not improve these capacities. SSCs displayed a greater capacity for survival and colonization in recipient seminiferous tubules; however, PB-MSCs were observed in all sections of testis after two weeks of transplantation. • MSCs are multipotent precursors with immune evasive capacity. • Bovine MSCs colonization capacity was evaluated in seminiferous tubules. • AT-MSCs were able to survive and colonize in allogenic recipient testis. • PB-MSCs were observed in all sections of testis after two weeks of transplantation. • SSCs had greater capacity for survival and colonization in recipient testis. [ABSTRACT FROM AUTHOR]

Anti-Müllerian hormone (AMH) has become a pivotal subject in the study of testicular descent, maturation, integrity, and male fertility. Recent studies explored its roles and implications across various domestic species. A prominent approach involved the understanding of the modulation of AMH in reproductive disorders, including cryptorchidism. While substantial findings have been reported in dogs, ruminants, swine, and horses, data on AMH in feline cryptorchidism remains limited. Here, we aimed to bridge this gap by comparing AMH serum levels among cryptorchid, healthy intact, and castrated tomcats, employing an enzyme-linked immunosorbent assay (ELISA) kit for quantification. In addition, AMH expression in retained and descended testes was evaluated through immunohistochemistry, with positive staining quantified via pixel analysis in two distinct regions of interest: the seminiferous tubule and the interstitial space. Furthermore, tissue samples were subjected to histological evaluation and morphometric analysis, which included the calculation of seminiferous tubule areas (STA) and assessment of Johnsen scores. Thus, the relationship between AMH expression, altered testicular histology, and impaired spermatogenesis could be examined. The expression of AMH in retained and descended testes, was investigated, and the relationship between AMH expression, altered testicular histology, and impaired spermatogenesis was examined. Mean serum AMH levels differed significantly (P < 0.001) across the different groups being 15.35 ± 4.66 ng/mL (mean ± SD) in healthy intact tomcats (n = 15), 25.55 ± 2.86 ng/mL (mean ± SD) in cryptorchids (n = 10) and below 0.015 ng/mL in castrated cats (n = 10). STAs and Johnsen scores were significantly reduced in retained testes when compared to descended gonads (P < 0.01). Furthermore, serum AMH was negatively correlated with both the STA (ρ = −0.725, P < 0.001) and the Johnsen scores (ρ = −0.699, P < 0.001), suggesting its potential value for tissue integrity and spermatogenesis evaluation. In addition, positive immunostaining was significantly higher in retained testes (P < 0.05), especially in the interstitial space (P < 0.01), suggesting an involvement of the Leydig cells. Additionally, the increased interstitial expression was linked to the degree of tissue degeneration and the impaired spermatogenesis being negatively corelated with both Johnsen scores (ρ = −0.309, P < 0.01) and STA (ρ = −0.208, P < 0.05). Our findings underscore the potential of AMH in assessing testicular health and reveal possible interspecific differences, stressing the need for further investigation in cats. • Cryptorchid tomcats have higher serum AMH levels than castrated or intact tomcats, but more research is needed to detect age-related thresholds. • Elevated AMH levels correlate with reduced seminiferous tubule area and lower Johnsen scores, indicating testicular degeneration. • Retained feline testicles show increased AMH in interstitial space, suggesting a species-specific expression pattern. • High AMH in retained testicles suggests increased Leydig cell receptor interaction, possibly altering steroidogenic activity. [ABSTRACT FROM AUTHOR]

Coronavirus disease 2019 (COVID-19) reportedly affects male reproductive function by causing spermatogenesis dysfunction and suppressing testosterone secretion. However, the relationship between COVID-19 and impaired reproductive function, such as whether these effects on reproductive function are a direct effect of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) infection in male reproductive organs or an indirect effect of high fever, is not known. Here, we examined whether the cell entry molecules of SARS-CoV-2, namely, ACE2, NRP1, TMPRSS2, and FURIN, are expressed in the male reproductive organs using the testes and accessory gonads of macaques during the breeding season. RT-PCR expression analysis showed that the testes alone expressed all four molecules. Immunohistochemical staining of testis tissue sections revealed that ACE2 is expressed in Leydig cells and the apical region of Sertoli cells, whereas NRP1 is expressed in the cell bodies surrounding the Leydig and Sertoli cell nuclei. FURIN is mainly expressed in Leydig cells, secondary spermatocytes, and spermatids. However, TMPRSS2 immunopositive cells were not observed. Therefore, it was not possible to observe cells expressing all four molecules in the gonads and accessory gonads of male primates. These results suggest that SARS-CoV-2 is unlikely to directly affect spermatogenesis in primates or proliferate in cells of the seminiferous tubules and undergo release into the semen through the previously known ACE2-mediated infection route. However, the expression of three molecules, including ACE2, was observed in Leydig cells, suggesting that testosterone synthesis and secretion may be affected when primates, including humans, are infected with SARS-CoV-2. [ABSTRACT FROM AUTHOR]

This research was designed to study the effect of silymarin dosage on the histological structure of the testicle in rabbits and the impact it has on the level of sperm production in these animals. Thirty adult male rabbits were used to complete this work. Their ages ranged between 5 to 6 months, while their weight ranged between 1,500 to 1,700 grams. The animals were divided into three groups based on the amount of dose of silymarin given as follows (C- 10 rabbits given free diet served as a control group: T1-10 rabbits treated with an oral daily dose of 0.5 ml per kg B.W. T2-10 rabbits treated with orally daily dose 1 ml per kg B.W.). The various types of cells were identified in this study based on several criteria, the most important of which are the external appearance of these cells during the series of divisions, the size of the cell, and the shape of the nucleus. The results showed an improvement in the architecture of seminiferous tubule epithelium, the disappearance of vacuolar degenerations in more than 95% of this epithelium, and an increase in the number of mature sperm in the center of each tubule in addition to increasing all parameters of Leydig cells. In conclusion, the testicles in rabbits exposed to silymarin are positively affected, leading to improved sperm production.

The testis serves as the primary site for spermatogenesis in mammals and is a crucial organ for the secretion of male hormones. Heat stress (HS) can have adverse effects on the seminiferous tubules, sperm quality, and sperm fertilization capability within the testis. Despite numerous previous studies describing various time points after heat stress in mice, a systematic and comprehensive dataset on heat stress and recovery in mice has been lacking. This study aimed to explore the gene expression changes in the recovery of multiple seminiferous epithelial cycles and spermatogenic cycles in mouse testicles after heat stress. We obtained high-throughput bulk RNA-seq data from testicular tissue of 4 NC mice and 32 HS mice (divided into 9 groups: NC, 30 min, 2 h, 6 h, 24 h, 3d, 8d, 24d, 47d, and 95d) and illustrated the dynamic changes in differential genes. This data set provides valuable insights into the detailed dynamic changes of one or more spermatogenic cycles after heat stress in mouse testicles, as well as the molecular mechanisms involved. [ABSTRACT FROM AUTHOR]

Bee-derived pharmaceutical products, including propolis (PRO) and royal jelly (ROJ), possess outstanding pharmacological properties. However, their efficiency in counteracting the deleterious influences of cadmium (Cd) in testes and the relevant mechanisms entail further investigations. Therefore, this study sheds light on the therapeutic efficacy of PRO and ROJ against testicular dysfunction and infertility induced by Cd. Toward this end, 30 mature male Wistar albino rats were randomly divided into six groups (5 animals/group), including (I) control, (II) Cd, (III) PRO, (IV) ROJ, (V) PRO + Cd, and (VI) ROJ + Cd groups. Furthermore, antioxidant factors, semen quality, hormonal levels, steroidogenic enzymes, and genotoxicity were assessed. Moreover, histopathological and ultrastructural attributes and offspring rates were investigated. The Cd-treated group revealed marked reductions in reduced glutathione (GSH), total antioxidant capacity (TAC), and superoxide dismutase (SOD) with an amplification of lipid peroxidation in testes, indicating disruption of the antioxidant defense system. Furthermore, myeloperoxidase (MPO) activity and DNA damage were significantly heightened, implying inflammation and genotoxicity, respectively. Moreover, steroidogenic enzymes, including 17β-Hydroxy Steroid Dehydrogenase 3 (HSD17b3), 3β-Hydroxy Steroid Dehydrogenase 2 (HSD3b2), 17α-hydroxylase/17,20-lyase (CYP17A1), and steroid 5α-reductase 2 (SRD5A2) were markedly diminished accompanied with disorders in luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone. Besides, spermatozoa quality was reduced, associated with a diminution in the diameter of seminiferous tubules. By contrast, PRO or ROJ significantly protected and/or counteracted the Cd-induced pathophysiological consequences, ameliorating antioxidant and inflammatory biomarkers, steroidogenic enzymes, hormonal levels, and sperm properties, along with lessening DNA impairments. Critically, histological and ultrastructural analyses manifested several anomalies in the testicular tissues of the Cd-administered group, including the Leydig and Sertoli cells and spermatozoa. Conversely, PRO or ROJ sustained testicular tissues' structure, enhancing spermatozoa integrity and productivity. Interestingly, treatment with PRO or ROJ improved fertility indices through offspring rates compared to the Cd-animal group. Our data suggest that PRO is a more effective countermeasure than ROJ against Cd toxicity for securing the delicate testicular microenvironment for spermatogenesis and steroidogenesis. [ABSTRACT FROM AUTHOR]

Background: Leprosy affects various organs in addition to skin, eyes, and peripheral nerves. Testicular involvement in leprosy patients is common and causes disturbance in endocrine function of the testis and results in hypogonadism. Hypogonadism is frequently undiagnosed and underreported. Objective: This study aimed to assess hypogonadism and associated factors among leprosy patients at Alert Comprehensive Specialized Hospital, Ethiopia. Methods: A cross-sectional study design was used in which consecutive 146 male leprosy patients aged between 18 to 65 years attending outpatient follow-up at leprosy outpatient clinic were included. Data was gathered both from patient charts and through patients' interviews. Androgen deficiency symptoms were assessed by androgen deficiency in the aging male questionnaire, and 5ml of blood samples were taken from study participants and serum total testosterone, LH, and FSH were analyzed by Electrochemiluminescence method. Statistical correlation was assessed by Spearman correlation. A multivariable binary logistic regression model was used to identify the independent factors associated with hypogonadism and P-value <0.05 was used to declare statistical significance. Results: The prevalence of hypogonadism was 39 (26.7%). Out of this, 34 (87.2%) had primary hypogonadism, whereas 5 (12.8%) had secondary hypogonadism. Total testosterone was inversely correlated with Body mass index (r = -0.37, p = 0.002), Luteinizing hormone (r = -0.43, p <0.001), and Follicular stimulating hormone (r = -0.42, p< 0.001). However, Total testosterone was not significantly correlated with age (r = -0.019, p = 0.81). BMI [AOR = 1.32, 95%CI (1.16–1.51)] and grade-II disability [AOR = 3.80, 95%CI (1.23–11.64)] were identified as independent risk factors for hypogonadism. Conclusion: Nearly one-fourth of male leprosy patients had hypogonadism. Overweight and grade-II disability were independent risk factors for hypogonadism. Author summary: Leprosy affects various organs in addition to skin, eyes and peripheral nerves. Testicular involvement in leprosy patients is common and causes disturbance in endocrine function of the testis and results in hypogonadism. Mycobacterium leprae reach the testicles through skin tissue invasion, blood or the lymphatic system, and the testes can serve as a reservoir for Mycobacterium leprae. A lower testicular temperature than internal body temperature may promote M. leprae growth in the testicle. It causes thickening and constriction of vessels, interstitial fibrosis, hyalinization and obliteration of seminiferous tubules and Leydig cells. These cause low testosterone productions among leprosy patients. The prevalence and risk factors of hypogonadism among male leprosy patients vary in different settings. In this study area the prevalence of hypogonadism was 26.7%. Overweight and grade-II disability were identified as independent risk factors for hypogonadism. [ABSTRACT FROM AUTHOR]

Azoospermia, the absence of sperm cells in semen, affects around 15% of infertile males. Sertoli cell-only syndrome (SCOS) is the most common pathological lesion in the background of non-obstructive azoospermia and is characterised by the complete absence of germinal epithelium, with Sertoli cells exclusively present in the seminiferous tubules. Studies have shown a correlation between successful spermatogenesis and male fertility with lipid composition of spermatozoa, semen, seminal plasma or testis. The aim of this research was to discover the correlation between the Johnsen scoring system and phospholipid expressions in testicular cryosections of SCOS patients. MALDI imaging mass spectrometry is used to determine spatial distributions of molecular species, such as phospholipids. Phosphatidylcholines (PCs), phosphatidylethanolamines (PEs) and sphingomyelins (SMs) are the most abundant phospholipids in mammalian cells and testis. SMs, the structural components of plasma membranes, are crucial for spermatogenesis and sperm function. Plasmalogens, are unique PCs in testis with strong antioxidative properties. This study, using imaging mass spectrometry, demonstrates the local distribution of phospholipids, particularly SMs, PCs, plasmalogens and PEs in human testicular samples with SCOS for the first time. This study found a strong relationship between the Johnsen scoring system and phospholipid expression levels in human testicular tissues. Future findings could enable routine diagnostic techniques during microTESE procedures for successful sperm extraction. [ABSTRACT FROM AUTHOR]

The present study describes the morphological and immunohistochemical characteristics of a case of diffuse seminoma in a 16‐year‐old male mixed‐breed horse. According to the owner, the animal's left testicle had been gradually increasing in size over a period of 2 months. On palpation, the testicle had a firm consistency, with no sensitivity to digital pressure, was adhered to the scrotum and measuring 16 cm × 8 cm. In the ultrasound examination, it presented a heterogeneous texture and areas of hypoechogenic echogenicity without visualization of the mediastinum. Therefore, the bilateral orchiectomy was performed. After the surgical procedure, it was found that the affected testicle presented a firm mass measuring 9 cm × 7 cm × 3.5 cm. Histologically, a multilobulated, non‐encapsulated and invasive tumour mass was found, which replaced the seminiferous tubules, consisting of polygonal cells arranged in a mantle that varied from cohesive to loosely cohesive, supported by a scarce fibrous stroma. In the immunohistochemical examination, the neoplastic cells showed positive immunolabelling for OCT4 and C‐KIT. In this report, the physical examination combined with the ultrasonographic examination were fundamental to the therapeutic management of the case, and the final diagnosis was made after histopathological and immunohistochemical tests. [ABSTRACT FROM AUTHOR]

Background: Plastic-based products are ubiquitous due to their tremendous utility in our daily lives. Nanoplastic (NP) and microplastic (MP) pollution has become a severe threat to the planet and is a growing concern. It has been widely reported that polystyrene (PS) MPs are severely toxic to the male reproduction system, with effects including decreased sperm parameters, impaired spermatogenesis, and damaged testicular structures. However, the molecular mechanisms for impaired spermatogenesis remain poorly understood. Methods: C57BL/6 male mice were treated with PS-NPs (80 nm) and PS-MPs (5 μm) by oral gavage every day for 60 days. A series of morphological analyses were completed to explore the influence of PS-NP and PS-MP exposure on the testes. Compared to other cell types in the seminiferous tubule, PS-NP and PS-MP exposure can lead to decreased spermatocytes. Then, more refined molecular typing was further performed based on gene expression profiles to better understand the common and specific molecular characteristics after exposure to PS-NPs and PS-MPs. Results: There were 1794 common DEGs across the PS-NP groups at three different doses and 1433 common DEGs across the PS-MP groups at three different doses. GO and KEGG analyses of the common DEGs in the PS-NP and PS-MP groups were performed to enrich the common and specific functional progress and signaling pathways, including 349 co-enriched GO entries and 13 co-enriched pathways. Moreover, 348 GO entries and 33 pathways were specifically enriched in the PS-NP group, while 526 GO entries and 15 pathways were specifically enriched in the PS-MPs group. Conclusions: PS-NPs were predominantly involved in regulating retinoic acid metabolism, whereas PS-MPs primarily influenced pyruvate metabolism and thyroid hormone metabolism. Our results highlight the different molecular mechanisms of PS-NPs and PS-MPs in the impairment of spermatogenesis in male mammals for the first time, providing valuable insights into the precise mechanisms of PS-NPs and PS-MPs in male reproduction. [ABSTRACT FROM AUTHOR]

Histological studies in the liver, lung and kidney of albino mice fed uninfested and infested wheat grains by granary weevil Sitophilus granaries were tested in the Agricultural Zoology laboratory of the Plant Protection Department, Faculty of Agriculture, Minia University, El-Minya, Egypt. Cross-section of the liver of an albino female fed healthy, uninfested grains, showed no bleeding or megakaryocytes. Sections in the liver of male albino mice fed on infested wheat grains by granary weevil S. granaries showed several megakaryocytes appeared in sinusoids as well as hemorrhage and congestion appeared in liver sections.Cross-section in the lungs of albino male mice fed healthy, uninfested grains, showed no focal chronic alveolitis. However histological studies on the lungs of albino mice fed infested wheat grains showed the thickness of alveolar walls with fibrosis, focal chronic alveolitis with fibrosis was likewise clear with a slight buildup of histiocytes around the terminal bronchiole and into the alveolar spaces.Bleeding inside bronchioles and outside alveoli also appeared. Moreover, kidney sections of male albino mice fed uninfested wheat grains by granary weevil showed normal kidney structure, while sections in kidneys of male albino mice fed infested wheat grains by granary weevil S. granariusshowed many distortions of the tubules whose cuboidal cells are “vacuolated” with the absence of a clear brush border (microvilli) of the proximal convoluted tubules. Hemorrhage appeared inside and outside seminiferous tubules. Thus, the gained data exposed significant changes in the liver, lung, and kidney of male albino mice fed infested wheat grains with S. granaries. [ABSTRACT FROM AUTHOR]

Pesticides like atrazine which are frequently present in everyday surroundings, have adverse impacts on human health and may contribute to male infertility. The work aimed to analyze the histological and biochemical effects of atrazine on the testis in adult albino rats and whether co-administration with resveratrol could reverse the effect of atrazine. Forty adult male albino rats in good health participated in this study. They were categorized at random into four groups: the Group Ӏ received water through a gastric tube for two months every day, the Group ӀӀ received resveratrol (20 mg/kg body weight (b.w.)) through a gastric tube for two months every day, the Group ӀӀӀ received atrazine (50 mg/kg bw) through a gastric tube for two months every day, the Group ӀV received concomitant doses of atrazine and resveratrol for two months every day. The testes of the animals were then carefully removed and prepared for biochemical, immunohistochemical, light, and electron microscopic studies. Atrazine exposure led to a significant decrease in serum testosterone hormone level, upregulation of caspase 3 and iNOS mRNA levels, destructed seminiferous tubules with few sperms in their lumens, many collagen fibres accumulation in the tunica albuginea and the interstitium, abnormal morphology of some sperms as well as many vacuolations, and damaged mitochondria in the cytoplasm of many germ cells. Concomitant administration of resveratrol can improve these adverse effects. It was concluded that atrazine exposure is toxic to the testis and impairs male fertility in adult rat and coadministration of resveratrol guards against this toxicity. [ABSTRACT FROM AUTHOR]

Metal toxicity from lead affects reproductive organ function by activating reactive oxygen species processes. This study aims to see how α-tocopherol and zinc sulfate (ZnSO4) affect gonads, liver, follicle-stimulating hormone, luteinizing hormone, spermatogenesis (the amount of spermatogonia, spermatocytes, and spermatids), and Leydig cells in male albino rats (Rattus norvegicus) exposed to lead acetate Pb(CH3COO)2. The samples used were 25 male Wistar rats aged 4 months, separated into five groups. All treatment groups were exposed to Pb(CH3COO)2 at a level of 50 mg/kg body weight (BW). The T1 group was given a dosage of 100 mg/kg BW of α-tocopherol. The ZnSO4 was given to the T2 group at a dose of 0.54 mg/kg BW. Meanwhile, the T3 group was given a mixture of ZnSO4 at 0.54 mg/kg BW and α-tocopherol at 100 mg/kg BW. All treatment is given orally for 30 days. ELISA test was carried out to determine the level of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in a blood plasma sample. Histopathological observations made on the liver included counting damaged cells and seminiferous tubules including counting the amount of spermatogonia, spermatocytes, spermatids, Sertoli cells, and Leydig cells. Using SPSS 20 software, the collected data were analyzed using analysis of variance, followed by Duncan's test with a 95% simultaneous confidence level. The highest average levels of FSH and LH in the T3 group were 3.6162 mIU/mL and 14.9658 mIU/mL. The finding showed that Pb(CH3COO)2 caused disruptions in the spermatogenesis and Leydig cell processes. Exogenous antioxidants in combination with ZnSO4 and α-tocopherol had a significant effect on enhancing reproductive performance in animals exposed to Pb(CH3COO)2. [ABSTRACT FROM AUTHOR]

Background: The goal of this study was to analyze the prophylactic effects of quercetin (Qu) against testes damage induced by 2600 MHz electromagnetic field (EMF) in rats. Materials and Methods: Thirty-two male rats (Wistar-Albino) were indiscriminately separated into four groups which were named: Control, Shamexposed, EMF-exposed (EMF, 1 h day-1 for 30 days), and EMF + Qu (100 mg/kg/daily) groups. After 30 days, the rats were sacrificed, testicular tissues were taken, and routine procedures were performed for histological and immunohistochemical evaluations. Results: When the testicular tissue of the control group was evaluated histopathologically, it was concluded that structures such as Leydig cells and seminiferous tubules were in normal condition and there was no change. No bleeding or inflammation was observed in the general structure of testicular tissues. Irregularities in spermatogenic cell configuration and shedding in the seminiferous epithelium were observed in the EMF and EMF + Qu groups. Findings close to our control group were detected in the sham-exposed and EMF + Qu groups. In addition, tumor necrosis factor-a (TNF-a) immune reactions in were EMF+ Qu group was decreased compared to the EMF group. Conclusion: In histopathological and immunohistochemical evaluation, it has been proven that EMF causes damage to testicular tissue. However, Qu demonstrated partial ameliorative effects on the pathological findings of the current study. [ABSTRACT FROM AUTHOR]

Duplicate testes lined in series were observed in the right scrotum of a 6-week-old Sprague-Dawley rat in a single-dose toxicity study. Of the two right testicles, one was spherical and less than half the size of a normal testis. The other was oval-shaped, slightly smaller than a normal testis, and possessed clear, tortuous blood vessels similar to those of a normal testis. Each right testis was grossly separated but faced the intertesticular adipose tissue and was sparsely joined by thin cord-like structures. Only one epididymis covered or encompassed the two right testes. The caput epididymis was attached to the smaller spherical testis, whereas the cauda epididymis was attached to the oval testis. Histopathological examination revealed that the smaller spherical testis on the right side and the testis on the left side were normal. The oval-shaped testis on the right exhibited markedly dilated degenerative seminiferous tubules with one to two layers of Sertoli or germ cells, and almost no spermatogenesis was observed. Multinucleated germ cells were observed in the lumen of the degenerated seminiferous tubules. The right epididymis was morphologically normal and contained few sperm in the epididymal duct of the tail. The cord-like structures between duplicate testes comprised fibrous and adipose tissues. Single efferent ductules, ectopic cartilage, and skeletal muscle tissues were buried in the adipose tissue. To our knowledge, this is the first report of spontaneous polyorchidism in a rodent. [ABSTRACT FROM AUTHOR]

Background: The lack of germ cells in the seminiferous tubules is a hallmark of Sertoli cell-only syndrome (SCOS), a leading cause of male infertility. Recently, stem cell therapy has gained attention as a possible method for treating this illness. Objectives: The aim of this study was to determine if stem cell treatment is effective for SCOS patients. Subjects and Methods: From 2020 to 2022, 87 males had been diagnosed with primary infertility owing to SCOS. The patients were evaluated in a thorough clinical manner, which included testing of their genes, hormone profiles, and testicular biopsies, among other things. The testes were injected with pure stem cells that had been taken from bone marrow. Semen analysis, hormone levels, and testicular volume were periodically evaluated over the 12-month follow-up period. Results: Following treatment, 69 patients (79.3% of the total) did not have sperm in their semen, whereas 18 patients (20.7% of the total) did. When compared to non-responders, respondents showed a statistically significant drop in FSH and LH levels and a rise in testosterone levels (P < 0.001). Hormonal changes were significantly different in the groups who responded and those that did not, and all responders had normal karyotyping and no AZF microdeletions. Conclusion: Hormonal improvements were seen in responding individuals, suggesting that stem cell therapy may be a viable option for treating SCOS. Nevertheless, a significant number of patients chose not to participate, underscoring the need for more studies to improve patient selection and treatment procedures. [ABSTRACT FROM AUTHOR]

BPA is a multifunctional endocrine disruptor with ubiquitous presence in aquatic ecosystems. The Mexican Central Plateau is an area severely impacted by pollution, inhabited by endemic viviparous fish. However, efforts to understand the effects of BPA on native species such as Goodea atripinnis are non-existent. This study focused on providing in vivo evidence of alterations in the testes of G. atripinnis males due to acute exposure to BPA at test concentrations of 1 mg/L, 10 mg/L, and 50 mg/L for 96 h. BPA exposition 1 mg/L and 10 mg/L showed degeneration and disorganization in germinal tissue. Furthermore, there was a notable decrease in sperm within the seminiferous tubules of males exposed to 10 mg/L of BPA. In all treatments, somatic cells had alterations by connective tissue thickening and an increase in collagen fibers. Additionally, inflammation and bleeding occurred in the testes of males exposed to 1 and 10 mg/L BPA. The alterations in the testes of G. atripinnis are related to BPA toxicity, which can lead to apoptosis in germ cells increasing connective tissue. Finally, even though the changes produced by BPA became evident in acute exposure (96 h), its effects are probably irreversible, compromising the reproduction of G. atripinnis. [ABSTRACT FROM AUTHOR]

Objectives: This study aims to comprehensively assess the therapeutic potential of clove extract in mitigating testicular damage and preserving male reproductive function under hypoxic conditions in male rats. Materials and Methods: We randomly divided 24 male rats into three groups: a hypoxia group that just received normal saline, a hypoxia + clove extract (4 mg/kg) group, and a sham operation group without treatment. After eight weeks, the serum level of oxidative stress parameters was examined, and the testicle histopathological examination, such as seminiferous tubule diameters and epithelium thickness, was assessed. Results: The serum malondialdehyde (MDA) concentration was significantly enhanced in the hypoxic group, and the levels of superoxide dismutase (SOD) and glutathione peroxidase (GPx) declined (P < 0.05). The diameters and thickness of the seminiferous tubule were diminished notably (P < 0.05). Johnson's score decreased in the hypoxic group. Treatment with clove extract also led to improvements in this parameter. Conclusions: Our study provides compelling evidence for the therapeutic potential of clove extract in mitigating testicular injury associated with varicocele-induced testicular hypoxia in male rats. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to investigate the potential protective impact of anthocyanin against tissue damage and oxidative stress provoked by varicocele within the testes of adult rats. Materials and Methods: A total of 32 male rats were divided into four groups as follows: a control group undergoing a sham procedure, a varicocele group without intervention (V), a varicocele group treated with anthocyanin (VA), and a group receiving anthocyanin treatment alone. Following a 56-day treatment period, the indicators of oxidative stress were gauged in the blood plasma, while histological modifications were evaluated utilizing the hematoxylin and eosin staining technique. Results: In the varicocele group treated with anthocyanin, we observed noteworthy enhancements in Johnsen score, epithelium thickness, and seminiferous tubule diameter compared to the untreated varicocele group. Treatment groups exhibited substantial elevations in testosterone levels and antioxidant enzyme levels. Furthermore, there was a reduction in the levels of malondialdehyde (MDA), an established marker of oxidative damage, and a decline in histological damage in the treatment groups. Conclusions: The outcomes underscore the potential safeguarding influence of anthocyanin against testicular damage stemming from varicocele induction, suggesting its beneficial role in countering oxidative stress and tissue impairment. [ABSTRACT FROM AUTHOR]

Context: Testicular regression syndrome (TRS), clinically termed "vanishing testes," is a condition with atrophy of testes in late fetal or early neonatal life. The spermatic cord ends blindly or into a small fibrovascular nubbin. Histopathology reveals a fibrovascular nodule with associated hemosiderin-laden macrophages and dystrophic calcification. Few cases show the presence of seminiferous tubules and germ cells. The presence of germ cells is associated with the risk of developing a testicular germ cell tumor. Aims: To study the histopathological features of testicular nubbins and to determine the presence of germ cells and associated neoplasm. Settings and Design: It is a retro-prospective observational study carried out on patients referred to a pediatric surgical pathology center in Miraj, who were clinically diagnosed with undescended testis, over a period of 1 year. Data were collected after the study was approved by the IEC-MGCH on December 19, 2022. Materials and Methods: Twenty patients diagnosed with cryptorchidism, undescended testis, or atrophic testis were included. Findings of hematoxylin and eosin-stained slides of each case were noted, and Prussian Blue (special staining) was performed in four cases. Statistical analysis used: As this is a descriptive observational study, simple arithmetic calculations were performed. No unique statistical analysis was performed. Results: Among twenty cases of vanishing testes syndrome, 20 cases showed fibrovascular tissue only, and 1 case showed the presence of seminiferous tubules with Sertoli cells. Conclusions: Vanishing testes syndrome is a rare condition that is difficult to diagnose on clinical examination, and a definitive diagnosis may be missed if the pathologist is unaware of the condition. Also, there is a risk of germ cell tumor if there are residual germ cells in the testicular nubbin. [ABSTRACT FROM AUTHOR]

Purpose: To determine the feasibility of high-frequency ultrasound (HFUS) for assessing seminiferous tubules and to understand high-resolution B-mode images of the testes in cases of azoospermia. Methods: We verified how the histopathological images of testicular biopsy specimens can be observed using HFUS images and measurement analysis of seminiferous tubules was performed to 28 testes of 14 cases with azoospermia who underwent preoperative ultrasound and microdissection testicular sperm extraction (micro-TESE). The population consisted of obstructive azoospermia (OA) and non-obstructive azoospermia (NOA), including Sertoli cell-only syndrome (SCOS), and the other pathologies. Statistical verification of differences in seminiferous tubule diameters among preoperative ultrasound examination, ultrasound examination of pathological specimens, and histopathological specimens. We also examined the imagingpathology correlation via a case series presentation, aiming to identify imaging markers of testicular pathology and determine the possibility of predicting each condition. Results: A comparison between HFUS images and histopathology from the same biopsy specimens suggested that ultrasonography could be seen as stereoscopic images due to its significantly greater slice thickness. The diameters of tubules were generally larger in pathological tissues as compared to ultrasonographic findings in OA and SCOS, but not in the other conditions. Comparisons provided insights into the predictability of SCOS and revealed imaging findings such as gaps between tubules and decreased diameter reflective of testicular damage. Conclusion: Seminiferous tubules can be observed however the diameter of seminiferous tubules varies in imaging and histopathology depending on the pathology. Imaging findings that reflect testicular damage and the predictability of SCOS were revealed in this study, but further verification is required. [ABSTRACT FROM AUTHOR]

Background: Camels are bred for their milk, meat, wool and hair, transportation, and their excrement as fuel. The seasonal reproduction of camel bull is accompanied by changes in sexual activity, the morphology, and function of the testes. This study aimed to evaluate the seasonal fluctuations in serum testosterone (T) levels as well as total antioxidant capacity (TAC) and malondialdehyde (MDA) levels in the testes of dromedary bulls (Camelus dromedarius) during the rutting and non-rutting seasons. Moreover, the impact of rutting season on the testicular size and histomorphology was also observed. Seventy mature dromedary bulls were divided into a rutting group (n = 35) and a non-rutting group (n = 35). From these bulls, blood samples and testes were collected during the rutting season (October to April) and non-rutting season (May to September) from a local slaughterhouse. Results: All parameters changed significantly during rutting and non-rutting periods in camel bulls. The levels of TAC in testes, and serum T were significantly (P < 0.05) higher in the rutting group than in the non-rutting group. However, testicular MDA was significantly (P < 0.05) lower in the rutting group than in the non-rutting group. TAC was negatively correlated with MDA (r = -0.59, p < 0.01). Moreover, in the rutting group and the non-rutting group, T was positively correlated with levels of TAC (r = 0.66, p < 0.0003). Additionally, testicular size (length, breadth, and thickness) was significantly greater in camels during the rutting season than in camels during the non-rutting season. Moreover, the number and diameter of seminiferous tubules, and spermatogenesis increased during the rutting season, whereas, the collagen content and apoptosis increased during the non-rutting season. Conclusion: This study revealed that the rutting normal breeding season (NBS, rutting group) was associated with higher levels of total antioxidant capacity (TAC), T, and spermatogenic activity while the collagen content, concentrations of MDA (the oxidative stress factor) and apoptosis (an outcome of oxidative stress) were lower than those in the low breeding season (LBS, non-rutting group). In addition, the testicular size and seminiferous tubule diameter and number were higher during the NBS. [ABSTRACT FROM AUTHOR]

Simple Summary: The detailed analysis of the structure of individual organs is necessary to understand the processes occurring within them. The complicated nature of male reproductive processes and the still insufficient knowledge of the structure of the male reproductive system prompted us to undertake the present research. In this study, using transmission electron microscopy (TEM) and histology staining, testes of sexually immature calves and reproductive bulls of the Polish Holstein-Friesian Black-and-White breed were analyzed. The obtained results were summarized and compared with previously published reports. The present manuscript presents complementary information on ultrastructure of seminiferous epithelium and interstitial tissue in cattle and provides an essential morphological background for further applied research. The purpose of this study was to analyze the ultrastructure of the testes of sexually immature calves and reproductive bulls of the Polish Holstein-Friesian Black-and-White breed. Utilizing TEM, this study identified three distinct stages of seminiferous tubule development in calves, characterized by varying shapes, distributions, and arrangements of individual cells. In immature animals, early developing spermatocytes, prespermatogonia, and pre-Sertoli cells were observed within the seminiferous tubules. In sexually mature bulls, all cells of the spermatogenic series were observed, situated on a thin, multilayered basal lamina, which forms characteristic undulations. An abundant smooth endoplasmic reticulum was observed in the cytoplasm of spermatogonia in both groups of animals, forming characteristic membranous swirls. In adult bulls, spermatogonia maintain contact with each other through numerous cytoplasmic bridges and cell connections, forming small spaces with visible microvilli between them. The ultrastructural analysis facilitated the identification of morphological changes occurring during the maturation of pre-Sertoli cells, transitioning from a large euchromatic nucleus to a nucleus in which the formation of characteristic vesicles and tubules could be observed. It should also be emphasized that two types of Sertoli cells, namely dark and light electron-dense cells, can be found in cattle. These cells differ from each other, indicating that they may perform different functions. The widespread recognition of the presence of two types of Sertoli cells in cattle will undoubtedly contribute to a better understanding of the processes occurring within the testes and provide a basis for further research in this area. [ABSTRACT FROM AUTHOR]

Background: The way of testicular tissue fixation directly affects the correlation and structural integrity between connective tissue and seminiferous tubules, which is essential for the study of male reproductive development. This study aimed to find the optimal fixative and fixation time to produce high-quality testicular histopathological sections, and provided a suitable foundation for in-depth study of male reproductive development with digital pathology technology. Methods: Testes were removed from both sides of 25 male C57BL/6 mice. Samples were fixed in three different fixatives, 10% neutral buffered formalin (10% NBF), modified Davidson's fluid (mDF), and Bouin's Fluid (BF), for 8, 12, and 24 h, respectively. Hematoxylin and eosin (H&E) staining, periodic acid Schiff-hematoxylin (PAS-h) staining, and immunohistochemistry (IHC) were used to evaluate the testicle morphology, staging of mouse seminiferous tubules, and protein preservation. Aperio ScanScope CS2 panoramic scanning was used to perform quantitative analyses. Results: H&E staining showed 10% NBF resulted in an approximately 15–17% reduction in the thickness of seminiferous epithelium. BF and mDF provided excellent results when staining acrosomes with PAS-h. IHC staining of synaptonemal complexes 3 (Sycp3) was superior in mDF compared to BF-fixed samples. Fixation in mDF and BF improved testis tissue morphology compared to 10% NBF. Conclusions: Quantitative analysis showed that BF exhibited a very low IHC staining efficiency and revealed that mouse testes fixed for 12 h with mDF, exhibited morphological details, excellent efficiency of PAS-h staining for seminiferous tubule staging, and IHC results. In addition, the morphological damage of testis was prolonged with the duration of fixation time. [ABSTRACT FROM AUTHOR]

Background: We recently reported that extract prepared from the aerial part of Cichorium intybus L. (CE) possesses hepatoprotective, hypolipidemic, and hypoglycemic properties. This paper focuses on the effects of CE on the male rat reproductive system and the effects of this treatment on pregnancy and offspring development. Methods: The experimental male rats received 100 mg/kg bw/day, 500 mg/kg bw/day, and 1000 mg/kg bw/day of CE orally for 60 consecutive days. Rats that received tap water were used as controls. After treatment, we evaluated the effects of CE on the male reproductive system, fertility, and offspring development. Results: For CE-treated male rats, there was a significant increase in the (1) diameter of seminiferous tubules, (2) spermatogenic index, (3) number of total and motile spermatozoa, and (4) testosterone levels. Additionally, there was a decrease in the pre- and post-implantation death of the embryos in the CE-treated group. All pups born from CE-treated males demonstrated normal development. Conclusions: CE treatment significantly improved male reproductive functions. No adverse effects on pregnancy and offspring development were observed when males were treated with CE. Further clinical evaluation of CE should lead to the development of a safe and effective phytodrug for treating male infertility. [ABSTRACT FROM AUTHOR]

The petit (pet) locus is associated with dwarfism, testicular anomalies, severe thymic hypoplasia, and high postnatal lethality, which are inherited in autosomal recessive mode of inheritance in rats with a Wistar strain genetic background. Linkage analysis localized the pet locus between 98.7 Mb and 101.2 Mb on rat chromosome 9. Nucleotide sequence analysis identified 2 bp deletion in exon 2 of the Thap4 gene as the causative mutation for pet. This deletion causes a frameshift and premature termination codon, resulting in a truncated THAP4 protein lacking approximately two-thirds of the C-terminal side. Thap4 is expressed in various organs, including the testis and thymus in rats. To elucidate the biological function of THAP4 in other species, we generated Thap4 knockout mice lacking exon 2 of the Thap4 gene through genome editing. Thap4 knockout mice also exhibited dwarfism and small testis but did not show high postnatal lethality. Thymus weights of adult Thap4 knockout male mice were significantly higher compared to wild-type male mice. Although Thap4 knockout male mice were fertile, their testis contained seminiferous tubules with spermatogenesis and degenerative seminiferous tubules lacking germ cells. Additionally, we observed vacuoles in seminiferous tubules, and clusters of cells in the lumen in seminiferous tubules in Thap4 knockout male mice. These results demonstrate that spontaneous mutation of Thap4 gene in rats and knockout of Thap4 gene in mice both cause dwarfism and testicular anomalies. Thap4 gene in rats and mice is essential for normal testicular development, maintaining spermatogenesis throughout the entire region of seminiferous tubules. [ABSTRACT FROM AUTHOR]